T315I RGQ PCR Kit

Selective and sensitive detection using ARMS and Scorpions technologies

A standardized assay with ready-to-use solutions

An easy workflow with the Rotor-Gene Q instrument

The T315I RGQ PCR Kit enables detection of the T315I mutation in the ABL gene (exon 6) using real-time PCR on the Rotor-Gene Q 5plex HRM Instrument. The kit provides all necessary reagents, optimized for rapid and sensitive detection of a low percentage of mutant DNA in a background of wild-type genomic DNA. ARMS PCR technology combined with the Scorpions detection technology provides high sensitivity and specificity for the detection of specific mutations.

For 24 reactions: Two vials of T315I Control Reaction Mix and one vial of T315I Reaction Mix, Positive Control, Taq DNA Polymerase, Water for NTC and Water for Sample Dilution

The T315I RGQ PCR Kit is intended for research use only. Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.

Principle

The T315I RGQ PCR Kit is a qualitative ready-to-use kit for the detection of the T315I mutation in the ABL gene (exon 6) using polymerase chain reaction (PCR) on the Rotor-Gene Q 5plex HRM instrument.

ARMS
Allele- or mutation-specific amplification is achieved by ARMS. Taq DNA polymerase is effective at distinguishing between a match and a mismatch at the 3' end of a PCR primer. Specific mutated sequences are selectively amplified, even in samples where the majority of the sequences do not carry the mutation. When the primer is fully matched, the amplification proceeds with full efficiency. When the 3' base is mismatched, only low-level background amplification occurs.

Scorpions
Detection of amplification is performed using Scorpions. Scorpions are bifunctional molecules containing a PCR primer covalently linked to a probe. The fluorophore in this probe interacts with a quencher, also incorporated into the probe, that reduces fluorescence. When the probe binds to the amplicon during PCR, the fluorophore and quencher become separated. This leads to an increase in fluorescence from the reaction tube.

Procedure

The T315I RGQ PCR Kit comprises a two-step procedure. In the first step, the control assay is performed to assess the total amplifiable T315I ABL DNA in a sample. In the second step, both the mutation and control assays are performed to determine the presence or absence of mutant DNA.

Applications

The T315I RGQ PCR Kit enables detection of the T315I mutation within the ABL gene (exon 6) in a background of genomic DNA.